B类Ⅰ型清道夫受体的高表达是结肠癌患者的有利预后因素

发布时间：2018-05-14 04:05

本文选题：B类Ⅰ型清道夫受体 + 结肠癌　；参考：《山东大学》2017年硕士论文

【摘要】：研究背景结肠癌是临床中发病率较高的疾病。近年来随着人们生活水平的改善,结肠癌的发病率也在逐年的升高。在近年的调查中发现在所有癌症患者中结肠癌的发病率居全世界第3位,并且女性患者普遍高于男性患者,平均每年有数十万的结肠癌患者死亡,因此对于结肠癌的研究和治疗具有重要的临床意义。流行病学显示,结肠癌的发生与高脂饮食,肥胖,饮酒,缺乏锻炼密切相关。既往研究表明,结肠癌的进展与血脂水平异常,特别是血浆高密度脂蛋白(High-density lipoprotein,HDL)的水平下降相关。B 类 Ⅰ 型清道夫受体(scavenger receptor class B type Ⅰ,SR-BⅠ)是第一个在分子水平上得到确证的高密度脂蛋白受体,体内SR-BⅠ表达相对广泛,主要表达于肝脏和肾上腺、卵巢、睾丸等固醇激素生成组织。SR-BⅠ是HDL中胆固醇及胆固醇酯转运的重要受体,参与体内胆固醇逆向转运过程。SR-BⅠ作为体内HDL的一种生理相关性受体,除了介导HDL抗炎、抗血栓、抗氧化等作用外,它还具有其它多种生物学作用,如在脓毒症、丙型肝炎病毒入侵、炎症、先天免疫、正常红细胞成熟和雌性生育等方面都有重要的调节作用。随后研究证实,SR-BⅠ在某些肿瘤细胞系中高表达,包括前列腺癌、肝癌、卵巢癌、乳腺癌、结肠癌、胰腺癌和鼻咽癌。然而,SR-BⅠ在结肠癌组织中的表达情况及其临床意义尚没有相关研究报道。在本课题中,我们首先应用生物信息学分析了脂质代谢相关基因在结肠癌患者与正常人之间的不同表达水平以及各基因与临床病理变量的关系,证实了SCARB1是参与胆固醇内化的5个基因中唯一在结肠癌患者中上调的基因。其次,我们利用组织芯片免疫组织化学技术检测了 SR-BI在90例结肠癌组织及其相应癌旁组织标本的表达情况,分析了 SR-BI的表达与患者各临床病理变量、预后的关系,证实SR-BI的表达虽与各临床病理变量无相关性,但是影响结肠癌预后的独立危险因素。目的1.应用生物信息学分析脂质代谢相关基因在结肠癌患者与正常人之间的不同表达水平以及各基因与临床病理变量的关系。2.采用含90例结肠癌组织及其相应癌旁组织标本的组织芯片检测SR-BI蛋白的表达情况。3.分析SR-BI的表达与结肠癌患者各临床病理变量之间的关系,并探讨SR-BI的表达状态与患者的预后关系,以期寻找新的结肠癌预后指标和潜在的治疗靶点。方法1.以癌症基因库(The Cancer Genome Altas,TCGA)提供的245例结肠癌患者及21例正常人的17组与脂质代谢相关基因信息为研究对象,分析脂质代谢相关基因在结肠癌患者与正常人之间的不同表达水平以及各基因与临床病理变量的关系。脂质代谢相关基因在结肠癌患者与正常人之间的不同表达水平采用T检验进行评估。各基因与临床病理变量的关系采用单因素方差分析进行评价。2.采用免疫组织化学染色的方法对购自上海芯超生物科技有限公司的含90例结肠癌组织和相对应的癌旁组织的芯片进行SR-BI表达的检测,并分析其与结肠癌癌的发生发展、临床病理特征及患者预后的关系。免疫组织化学SR-BI染色的评分由两个独立的病理学家进行,屏蔽患者的临床信息和预后情况。评分的方法应用改进的免疫反应评分法(immune response scoring method,IRS),主要评价染色细胞的百分比(无染色,0;10%,1;10-50%,2;51-80%,3;80%,4)和染色强度(无染色,0;弱,1;中度,2;强,3)。染色强度和染色阳性细胞数比例的乘积得分(范围从0到12)作为每例患者的最终得分。在随后的统计分析中应用两级分层。肿瘤标本评分最后在0-4之间认为是SR-BI低表达,而≥6认为是高表达。SR-BI的表达与临床病理特征之间的关系采用卡方检验或Fisher精确检验进行评估,取决于不同的临床病理参数的数值性质和正态分布性。生存分析采用Kaplan-Meier生存分析进行评价,并用Log-rank检验各变量对生存时间(overall survival,OS)的影响。多因素分析采用Cox比例风险模型来计算,计算风险比(hazard ratio,HR)和95%可信区间(confidence interval,CI)。结果1.相比于正常人,5个基因在结肠癌患者中明显上调,SCARB1(P = 0.000),金属蛋白酶组织抑制物1(TIMP1,P = 0.000),基质金属蛋白酶9(MMP-9,P = 0.000),固醇调节元件结合转录因子1(SREBF1,P = 0.028),脂肪酸合酶(FASN,P = 0.000)。9个基因在结肠癌患者中明显下调,组织蛋白酶S(CTSS,P = 0.000),胆固醇脂化微囊蛋白1(CAV1,P = 0.001),极低密度脂蛋白受体相关蛋白(VLDLR,P = 0.000),ATP结合盒转运子1(ABCA1,P = 0.000),乙酰辅酶A乙酰转移酶1(ACAT1,P = 0.000),分化抗原簇36(CD36,P = 0.000),3-羟基-3-甲基-戊二酸-辅酶 A 还原酶(HMG-CoAR,P= 0.047),固醇调节元件结合转录因子2(SREBF2,=0.001),肝X受体激动剂 α(LXR-α,P = 0.000)。2.CD36(P = 0.002),CTSS(P = 0.028),VLDLR(P = 0.020)在结肠癌患者中的表达水平与淋巴结转移的阳性率相关。CTSS(P = 0.025),ACAT1(P=0.006),MMP-9(P=0.003),HMG-CoAR(P = 0.005)与 T 分期相关。LDLRP(P = 0.041)和MMP-9(P = 0.047)与肿瘤分级相关。值得注意的是SCARB1的表达水平与各临床病理变量无关。3.在90例结肠癌组织标本中,51例(56.7%)为SR-BⅠ高表达,39例(43.3%)为SR-BⅠ低表达。在90例相对应的癌旁组织中,48例(53.3%)为SR-BⅠ高表达,42例(46.7%)为SR-BⅠ低表达。SR-BⅠ在大多数结肠癌组织比相对应的癌旁组织表达高(P = 0.000)。4.与SCARB1的表达水平与各临床病理变量无关的结果一致,SR-BⅠ的表达水平与性别、年龄、肿瘤pTNM分期、肿瘤大小、肿瘤分级、淋巴结总数、淋巴结转移的阳性率、AJCC临床分期无关。5.在结肠癌组织中,SR-BⅠ低表达的患者比SR-BⅠ高表达的患者总体生存率(OS)缩短(log-rank检验,P = 0.042)。Cox回归多因素分析提示SR-BI的高表达是影响预后的独立有利因素,HR为0.502(P = 0.039),除了 SR-BI之外,淋巴结转移的阳性率也是结肠癌患者预后的独立因素(P = 0.007)。结论1.基因SCARB1的表达水平在结肠癌患者中上调。2.结肠癌组织SR-BI表达水平比癌旁组织高。3.结肠癌组织中SR-BI的表达水平与结肠癌患者的生存相关。SR-BI表达水平越低,预后越差。4.结肠癌组织中SR-BI的表达水平是结肠癌患者的独立预后因素。研究意义SR-BI的低表达状态与结肠癌患者不良预后有关,SR-BI的表达可能被作为结肠癌的恶性指标之一而应用于临床检测中。
[Abstract]:Background colon cancer is a disease with high incidence in clinic. In recent years, with the improvement of people's living standard, the incidence of colon cancer is also increasing year by year. In recent years, the incidence of colon cancer in all cancer patients is third in the world, and the female patients are generally higher than those of male patients. One hundred thousand of the colon cancer patients died, so the research and treatment of colon cancer has important clinical significance. Epidemiology shows that the occurrence of colon cancer is closely related to high fat diet, obesity, drinking and lack of exercise. Previous studies have shown that the progression of colon cancer and blood lipid levels are abnormal, especially the plasma high density lipoprotein (High-density lip). The level decline of oprotein, HDL) related.B class type I scavenger receptor (scavenger receptor class B type I, SR-B I) is the first high density lipoprotein receptor confirmed at the molecular level, and the expression of SR-B I in the body is relatively extensive, mainly expressed in the liver and adrenal, ovary, testis and other steroid hormone producing tissue.SR-B I is HDL .SR-B I, an important receptor for the transport of cholesterol and cholesteryl ester, is involved in the reverse transport of cholesterol in the body as a physiological receptor in the body of HDL. In addition to mediating the anti-inflammatory, antithrombotic and antioxidant functions of HDL, it also has many other biological effects, such as sepsis, hepatitis C virus invasion, inflammation, and innate immunity. SR-B I was highly expressed in some tumor cell lines, including prostate cancer, liver cancer, ovarian cancer, breast cancer, colon cancer, pancreatic cancer, and nasopharyngeal carcinoma. However, there is no correlation between the expression of SR-B I in colon cancer and its clinical significance. In this study, we first applied bioinformatics to analyze the different expression levels of lipid metabolism related genes in colon cancer patients and normal people, and the relationship between the genes and the clinicopathological variables, and confirmed that SCARB1 is the only gene for the 5 genes involved in cholesterol internalization in colon cancer patients. At the same time, we detected the expression of SR-BI in 90 cases of colon cancer tissue and its adjacent tissues by tissue microarray immunohistochemical technique, analyzed the relationship between the expression of SR-BI and the clinicopathological variables and the prognosis of the patients, and confirmed that the expression of SR-BI was not related to the clinicopathological variables, but it affected the prognosis of colon cancer. Independent risk factors. Objective 1. application of bioinformatics to analyze the different expression levels of lipid metabolism related genes in patients with colon cancer and normal people and the relationship between the genes and the clinicopathological variables.2. use the tissue core of 90 cases of colon cancer tissue and its corresponding paracancerous tissue samples to detect the expression of SR-BI protein.3. analysis S The relationship between the expression of R-BI and the clinicopathological variables of colon cancer patients, and the relationship between the expression of SR-BI and the prognosis of the patients, in order to find a new prognostic indicator and potential therapeutic targets for colon cancer. Method 1. the 17 groups of colon cancer patients and 21 normal persons were provided with the The Cancer Genome Altas (TCGA). The different expression levels of lipid metabolism related genes in colon cancer patients and normal people and the relationship between genes and clinicopathological variables are analyzed with lipid metabolism related gene information. The different expression levels of lipid metabolism related genes in colon cancer patients and normal people are evaluated by T test. The relationship with the clinicopathological variables was evaluated by single factor ANOVA..2. was used to detect the expression of SR-BI in 90 cases of colon cancer tissue and corresponding para cancerous tissues purchased from Shanghai core superbiological science and Technology Co., Ltd., and analyzed the occurrence and development of carcinoma of colon and clinical disease. The relationship between physical characteristics and patient prognosis. The score of immunohistochemical SR-BI staining was carried out by two independent pathologists to shield the patient's clinical information and prognosis. The scoring method was used to evaluate the percentage of dyed cells (no staining, 0; 10%, 1; 10-5) by the improved immune response score (Immune response scoring method, IRS). 0%, 2; 51-80%, 3; 80%, 4) and the intensity of dyed (no dyeing, 0; weak, 1; moderate, 2; 3). The product score (range from 0 to 12) was the final score of each patient (range from 0 to 12). The relationship between the expression of high expression of.SR-BI and the clinicopathological features was evaluated by chi square test or Fisher accurate test, depending on the numerical and normal distribution of different clinicopathological parameters. The survival analysis was evaluated by Kaplan-Meier survival analysis, and the Log-rank test was used to test the survival time (overall surviv). The effect of Al, OS). The multifactor analysis uses the Cox proportional hazard model to calculate the risk ratio (hazard ratio, HR) and the 95% confidence interval (confidence interval, CI). Results 1. compared to normal people, 5 genes were significantly up-regulated in colon cancer patients, SCARB1 (P = 0), metalloproteinase tissue inhibitor 1 (TIMP1, 0), and matrix metalloproteinases. Enzyme 9 (MMP-9, P = 0), sterol regulatory element combined with transcription factor 1 (SREBF1, P = 0.028), fatty acid synthase (FASN, P = 0).9 genes were significantly down regulated in colon cancer patients, cathepsin S (CTSS, P = 0), cholesterol lipoprotein microcystin 1 (CAV1, P = 0.001), extremely low density lipoprotein receptor related protein (0 = 0), binding Box transporter 1 (ABCA1, P = 0), acetyl coenzyme A acetyltransferase 1 (ACAT1, P = 0), differentiated antigen cluster 36 (CD36, P = 0), 3- hydroxyl -3- methyl - glutaric acid - coenzyme A reductase (HMG-CoAR, P= 0.047), sterol regulator element binding transcription factor 2. 0.028), the expression level of VLDLR (P = 0.020) in colon cancer patients was related to the positive rate of lymph node metastasis (.CTSS (P = 0.025), ACAT1 (P=0.006), MMP-9 (P=0.003), HMG-CoAR (P = 0.005) and T staging related.LDLRP (0.041) and P = 0.047). Of the 90 cases of colon cancer, 51 cases (56.7%) were high expression of SR-B I, 39 (43.3%) was SR-B I low expression. In the corresponding para cancerous tissues of 90 cases, 48 cases (53.3%) were high expression of SR-B I, 42 (46.7%) was SR-B I low expression.SR-B I expressed high (P = 0).4. and SCA in most colon cancer tissues than corresponding cancerous tissues. The expression level of RB1 was consistent with that of various clinicopathological variables. The expression level of SR-B I was not related to sex, age, tumor pTNM staging, tumor size, tumor classification, lymph node number, positive rate of lymph node metastasis, and.5. in colon cancer tissue, and the patients with low expression of SR-B I were higher than those with high expression of SR-B I. The survival rate (OS) shortening (log-rank test, P = 0.042).Cox regression multivariate analysis suggested that the high expression of SR-BI was an independent factor affecting the prognosis, HR was 0.502 (P = 0.039). Besides SR-BI, the positive rate of lymph node metastasis was also an independent factor (P = 0.007) for the prognosis of colon cancer patients. Conclusion the expression level of the 1. gene SCARB1 is in colon cancer patients. The expression level of SR-BI in the.2. colon cancer tissue is higher than that of the high.3. colon cancer tissue of the paracancerous tissue. The lower the expression level of SR-BI in the colon cancer patients, the lower the.SR-BI expression level, the worse the prognosis in.4. colon cancer tissue, the expression level of SR-BI is the independent preconditioning factor of colon cancer patients. The poor prognosis of patients with colorectal cancer is related. The expression of SR-BI may be used as a malignant marker of colon cancer and used in clinical detection.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.35

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